杨梅基因组DNA提取方法筛选和优化研究

选择具有代表性的杨梅品种20份,分别采用CTAB法和SDS法,选取嫩叶进行基因组DNA提取比较研究,结果表明,由于叶片中含有较高的酚类物质,相对来说CTAB法优于SDS法,提取的基因组DNA色泽白亮且数量多,而且同一提取方法如CTAB法,不同品种之间提取得率也有较大差异。在CTAB法的基础上,选取两个品种研究水浴时间对DNA提取的影响,发现DNA得率随水浴时间的延长呈现抛物线态势,最佳水浴时间随样品颗粒的大小而变化,一般最佳时间为30～40min,样品颗粒较细时水浴时间以30min为最佳,较粗时则为40min。添加异丙醇量以2/3为好,析出DNA的最佳离心速度和时间分别为5 000r/min和10～15min。

Optimization of Extraction Conditions and Methods for Genome DNA in Myrica

Both CTAB and SDS were compared for abstracting genome DNA using young leaves of 20 Myrica rubra varieties.The result showed that genome DNA for CTAB were more and blighter than for SDS because of upper hydroxybenzene content in the leaves of waxberry.Besides there were differences for extraction contents of genome DNA among varieties even though the same extraction method such as CTAB.It was studied effects of bath times on genone DNA quality in two varieties of waxberry Based on method CTAB,and indicated that genome DNA contents presented parabola state under different bath time.In general,the best bath time was 30~40 minutes and changed with size of sample leaves,and it was 30 min.for bit leaves while it was 40min.for massive ones.It was better for adding 2/3 isopropyl alcohol than 3/3 one.And the best centrifugal speed and time was 5 000r/min and 10~15min.,respectively.